Use

ABSTRACT

The present invention is relating to a new use of an immunosuppressant, for increasing an antifungal activity of a lipopeptide compound.

TECHNICAL FIELD

[0001] This invention relates to a new use of an immunosuppressant,which is useful in a medical field.

BACKGROUND ART

[0002] Various immunosuppressants are well known. And, a certainlipopeptide compound, and its related compounds are known to have anantifungal activity and inhibit an activity of β-1,3-glucan synthase(WO96/11210 and WO99/40108).

DISCLOSURE OF INVENTION

[0003] This invention relates to a new use of an immunosuppressant, forincreasing an antifungal activity of a lipopeptide compound.

[0004] Therefore, one object of the present invention is to provide anew use of an immunosuppressant for increasing an antifungal activity ofa lipopeptide compound.

[0005] Another object of this invention is to provide a method forincreasing an antifungal activity of a lipopeptide compound byadministering an effective amount of an immunosuppressant.

[0006] A further object of this invention is to provide a use of animmunosuppressant for manufacturing a medicament for increasing anantifungal activity of a lipopeptide compound.

[0007] Still further object of this invention is to provide acomposition comprising an immunosuppressant, for increasing anantifungal activity of a lipopeptide compound.

[0008] Preferable “immunosuppressant” is, for example, the tricyclicmacrolide shown in EP-0184162, WO89/05303, WO93/05058, WO96/31514, andso on, the disclosure of which is incorporated herein by reference.

[0009] As a particular example of the tricyclic macrolides compounds,the tricyclic compound of the following formula (II) can be exemplified.

[0010] (wherein each of adjacent pairs of R¹ and R² R³ and R⁴, and R⁵and R⁶ independently

[0011] (a) is two adjacent hydrogen atoms, but R² may also be an alkylgroup or

[0012] (b) may form another bond formed between the carbon atoms towhich they are attached;

[0013] R⁷ is a hydrogen atom, a hydroxy group, a protected hydroxygroup, or an alkoxy group, or an oxo group together with R¹;

[0014] R⁸ and R⁹ are independently a hydrogen atom or a hydroxy group;

[0015] R¹⁰ is a hydrogen atom, an alkyl group, an alkyl groupsubstituted by one or more hydroxy groups, an alkenyl group, an alkenylgroup substituted by one or more hydroxy groups, or an alkyl groupsubstituted by an oxo group;

[0016] X is an oxo group, (a hydrogen atom and a hydroxy group), (ahydrogen atom and a hydrogen atom), or a group represented by theformula —CH₂O—;

[0017] Y is an oxo group, (a hydrogen atom and a hydroxy group), (ahydrogen atom and a hydrogen atom), or a group represented by theformula N—NR¹¹R¹² or N—OR¹³;

[0018] R¹¹ and R¹² are independently a hydrogen atom, an alkyl group, anaryl group or a tosyl group;

[0019] R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and R²³ are independentlya hydrogen atom or an alkyl group;

[0020] R²⁴ is an optionally substituted ring system which may containone or more heteroatoms;

[0021] n is an integer of 1 or 2; and

[0022] in addition to the above definitions, Y, R¹⁰ and R²³, togetherwith the carbon atoms to which they are attached, may represent asaturated or unsaturated 5- or 6-membered nitrogen, sulfur and/or oxygencontaining heterocyclic ring optionally substituted by one or moregroups selected from the group consisting of an alkyl, a hydroxy, analkoxy, a benzyl, a group of the formula —CH₂Se(C₆H₅), and an alkylsubstituted by one or more hydroxy groups.

[0023] The definitions used in the above general formula (II) and thespecific and preferred examples thereof are now explained and set forthin detail.

[0024] The term “lower” means, unless otherwise indicated, a grouphaving 1 to 6 carbon atoms.

[0025] Preferable examples of the “alkyl groups” and an alkyl moiety ofthe “alkoxy group” include a straight or branched chain aliphatichydrocarbon residue, for example, a lower alkyl group such as methyl,ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, neopentyl and hexyl.

[0026] Preferable examples of the “alkenyl groups” include a straight orbranched chain aliphatic hydrocarbon residue having one double-bond, forexample, a lower alkenyl group such as vinyl, propenyl (e.g., allylgroup), butenyl, methylpropenyl, pentenyl and hexenyl.

[0027] Preferable examples of the “aryl groups” include phenyl, tolyl,xylyl, cumenyl, mesityl and naphthyl.

[0028] Preferable protective groups in the “protected hydroxy groups”and the “protected amino” are 1-(lower alkylthio)-(lower)alkyl groupsuch as a lower alkylthiomethyl group (e.g., methylthiomethyl,ethylthiomethyl, propylthiomethyl, isopropylthiomethyl, butylthiomethyl,isobutylthiomethyl, hexylthiomethyl, etc.), more preferably C₁-C₄alkylthiomethyl group, most preferably methylthiomethyl group;

[0029] trisubstituted silyl group such as a tri(lower)alkylsilyl (e.g.,trimethylsilyl, triethylsilyl, tributylsilyl, tert-butyldimethylsilyl,tri-tert-butylsilyl, etc.) or lower alkyl-diarylsilyl (e.g.,methyldiphenylsilyl, ethyldiphenylsilyl, propyldiphenylsilyl,tert-butyldiphenyl-silyl, etc.), more preferably tri(C₁-C₄)alkylsilylgroup and C₁-C₄ alkyldiphenylsilyl group, most preferablytert-butyldimethylsilyl group and tert-butyldiphenylsilyl group; and anacyl group such as an aliphatic, aromatic acyl group or an aliphaticacyl group substituted by an aromatic group, which are derived from acarboxylic acid, sulfonic acid or carbamic acid.

[0030] Examples of the aliphatic acyl groups include a lower alkanoylgroup optionally having one or more suitable substituents such ascarboxy, e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl,isovaleryl, pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl,carboxybutyryl, carboxyhexanoyl, etc.; acyclo(lower)alkoxy(lower)alkanoyl group optionally having one or moresuitable substituents such as lower alkyl, e.g., cyclopropyloxyacetyl,cyclobutyloxypropionyl, cycloheptyloxybutyryl, menthyloxyacetyl,menthyloxypropionyl, menthyloxybutyryl, menthyloxypentanoyl,menthyloxyhexanoyl, etc.; a camphorsulfonyl group; or a loweralkylcarbamoyl group having one or more suitable substituents such ascarboxy or protected carboxy, for example, carboxy(lower)alkylcarbamoylgroup (e.g., carboxymethylcarbamoyl, carboxyethylcarbamoyl,carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentylcarbamoyl,carboxyhexylcarbamoyl, etc.),tri-(lower)alkylsilyl(lower)alkoxycarbonyl(lower)alkylcarbamoyl group(e.g., trimethylsilylmethoxycarbonylethylcarbamoyl,trimethylsilylethoxycarbonylpropylcarbamoyl,triethylsilylethoxycarbonylpropylcarbamoyl,tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl,tri-methylsilylpropoxycarbonylbutylcarbamoyl, etc.) and so on.

[0031] Examples of the aromatic acyl groups include an aroyl groupoptionally having one or more suitable substituents such as nitro, e.g.,benzoyl, toluoyl, xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl,nitronaphthoyl, etc.; and an arenesulfonyl group optionally having oneor more suitable substituents such as halogen, e.g., benzenesulfonyl,toluenesulfonyl, xylenesulfonyl, naphthalenesulfonyl,fluorobenzenesulfonyl, chlorobenzenesulfonyl, bromobenzenesulfonyl,iodobenzenesulfonyl, etc.

[0032] Examples of the aliphatic acyl groups substituted by an aromaticgroup include ar(lower)alkanoyl group optionally having one or moresuitable substituents such as lower alkoxy or trihalo(lower)alkyl, e.g.,phenylacetyl, phenylpropionyl, phenylbutyryl,2-trifluoromethyl-2-methoxy-2-phenylacetyl,2-ethyl-2-trifluoromethyl-2-phenylacetyl,2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc.

[0033] More preferable acyl groups among the aforesaid acyl groups areC₁-C₄ alkanoyl group optionally having carboxy,cyclo(C₅-C₆)alkoxy(C₁-C₄)alkanoyl group having two (C₁-C₄) alkyls at thecycloalkyl moiety, camphorsulfonyl group, carboxy-(C₁-C₄)alkylcarbamoylgroup, tri (C₁-C₄) alkylsilyl (C₁-C₄) alkoxycarbonyl(C₁-C₄)-alkylcarbamoyl group, benzoyl group optionally having one or twonitro groups, benzenesulfonyl group having halogen, orphenyl(C₁-C₄)alkanoyl group having C₁-C₄ alkoxy and trihalo(C₁-C₄) alkylgroup. Among these, the most preferable ones are acetyl,carboxypropionyl, menthyloxyacetyl, camphorsulfonyl, benzoyl,nitrobenzoyl, dinitrobenzoyl, iodobenzenesulfonyl and2-trifluoromethyl-2-methoxy-2-phenylacetyl.

[0034] Preferable examples of the “5- or 6-membered nitrogen, sulfurand/or oxygen containing heterocyclic ring” include a pyrrolyl group anda tetrahydrofuryl group.

[0035] R²⁴ is an optionally substituted ring system which may containone or more heteroatoms, Preferable R²⁴ may be cyclo (C₅₋₇) alkyl groupoptionally having suitable substituents, and the following ones can beexemplified.

[0036] (a) a 3,4-di-oxo-cyclohexyl group;

[0037] (b) a 3-R²⁰-4-R²¹-cyclohexyl group,

[0038] in which

[0039] R²⁰ is hydroxy, an alkoxy group, an oxo group, or a—OCH₂OCH₂CH₂OCH₃ group, and

[0040] R²¹ is hydroxy, —OCN, an alkoxy group, a heteroaryloxy which maybe substituted by suitable substituents, 1- or 2-tetrazolyl, a—OCH₂OCH₂CH₂OCH₃ group, a protected hydroxy group, chloro, bromo, iodo,aminooxalyloxy, an azido group, p-tolyloxythiocarbonyloxy, orR²⁵R²⁶CHCOO—,

[0041] in which

[0042] R²⁵ is optionally protected hydroxy or protected amino, and

[0043] R²⁶ is hydrogen or methyl, or

[0044]  R²⁰ and R²¹ together form an oxygen atom in an epoxide ring; or

[0045] (c) cyclopentyl group substituted by methoxymethyl, optionallyprotected hydroxymethyl, acyloxymethyl

[0046] (in which the acyl moiety optionally contains either adimethylamino group which may be quaternized, or a carboxy group whichmay be esterified), one or more amino and/or hydroxy groups which may beprotected, or aminooxalyloxymethyl. A preferred example is a2-formyl-cyclopentyl group.

[0047] “A heteroaryl which may be substituted by suitable substituents”moiety of the “heteroaryloxy which may be substituted by suitablesubstituents” may be the ones exemplified for R¹ of the compound of theformula of EP-A-532,088, with preference given to1-hydroxyethylindol-5-yl, the disclosure of which is incorporated hereinby reference.

[0048] The tricyclic compounds (II) and its pharmaceutically acceptablesalt for use in accordance with this invention are well known to haveexcellent immunosuppressive activity, antimicrobial activity and otherpharmacological activities and, as such, be of value for the treatmentor prevention of rejection reactions by transplantation of organs ortissues, graft-vs-host diseases, autoimmune diseases, and infectiousdiseases [EP-A-0184162, EP-A-0323042, EP-A-423714, EP-A-427680,EP-A-465426, EP-A-480623, EP-A-532088, EP-A-532089, EP-A-569337,EP-A-626385, WO89/05303, WO93/05058, WO96/31514, WO91/13889, WO91/19495,WO93/04680, WO93/5059, etc.], the disclosures of which are incorporatedherein by reference.

[0049] Particularly, the compounds which are designated as FR900506(=FK506), FR900520 (ascomycin), FR900523, and FR900525 are productsproduced by microorganisms of the genus Streptomyces, such asStreptomyces tsukubaensis No. 9993 [deposited with National Institute ofBioscience and Human Technology Agency of Industrial Science andTechnology (formerly Fermentation Research Institute Agency ofIndustrial Science and Technology ), at 1-3, Higashi 1-chome,Tsukuba-shi, Ibaraki, Japan, date of deposit Oct. 5, 1984, accessionnumber FERM BP-927] or Streptomyces hygroscopicus subsp. yakushimaensisNo. 7238 [deposited with National Institute of Bioscience and HumanTechnology Agency of Industrial Science and Technology (formerlyFermentation Research Institute Agency of Industrial Science andTechnology ), at 1-3, Higashi 1-chome, Tsukuba-shi, Ibaraki, Japan, dateof deposit Jan. 12, 1985, accession number FERM BP-928][EP-A-0184162].The FK506 (general name: tacrolimus) of the following chemical formula,in particular, is a representative compound.

[0050] Chemical name:17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone

[0051] The preferred examples of the tricyclic compounds (II) are theones, wherein each of adjacent pairs of R³ and R⁴ or Rs and R⁶independently form another bond formed between the carbon atoms to whichthey are attached;

[0052] each of R⁸ and R²³ is independently a hydrogen atom;

[0053] R⁹ is a hydroxy group;

[0054] R¹⁰ is a methyl group, an ethyl group, a propyl group or an allylgroup;

[0055] X is (a hydrogen atom and a hydrogen atom) or an oxo group;

[0056] Y is an oxo group;

[0057] each of R¹⁴ R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, and R²² is a methyl group;

[0058] R²⁴ is a 3-R²⁰-4-R²¹-cyclohexyl group,

[0059] in which

[0060] R²⁰ is hydroxy, an alkoxy group, an oxo group, or a—OCH₂OCH₂CH₂OCH₃ group, and

[0061] R²¹ is hydroxy, —OCN, an alkoxy group, a heteroaryloxy which maybe substituted by suitable substituents, 1- or 2-tetrazolyl, a—OCH₂OCH₂CH₂OCH₃ group, a protected hydroxy group, chloro, bromo, iodo,aminooxalyloxy, an azido group, p-tolyloxythiocarbonyloxy, orR²⁵R²⁶CHOO—

[0062] in which

[0063] R²⁵ is optionally protected hydroxy or protected amino, and

[0064] R²⁶ is hydrogen or methyl, or

[0065]  R²⁰ and R²¹ together form an oxygen atom in an epoxide ring; and

[0066] n is an integer of 1 or 2.

[0067] The most preferable tricyclic compounds (II) is, in addition toFK506, ascomycin derivatives such as halogenated-ascomycin (e.g.,33-epi-chloro-33-desoxyascomycin), which is disclosed in EP 427,680,example 66a.

[0068] As the other preferable example of the immunosuppressant,rapamycin [THE MERCK INDEX (12th edition), No. 8288] and its derivativescan be exemplified. Preferred example of the derivatives is anO-substituted derivative in which the hydroxy in position 40 of formulaA illustrated at page 1 of WO 95/16691, incorporated herein byreference, is replaced by —OR₁ in which R₁ is hydroxyalkyl,hydroalkoxyalkyl, acylaminoalkyl and aminoalkyl; for example40-O-(2-hydroxy)ethyl-rapamycin, 40-O-(3-hydroxy)propyl-rapamycin,40-O-[2-(2-hydroxy)ethoxy]ethyl-rapamycin and40-O-(2-acetaminoethyl)-rapamycin. These O-substituted derivatives maybe produced by reacting rapamycin (or dihydro or deoxo-rapamycin) withan organic radical attached to a leaving group (for example RX where Ris the organic radical which is desired as the O-substituent, such as analkyl, allyl, or benzyl moiety, and X is a leaving group such asCCl₃C(NH)O or CF₃SO₃) under suitable reaction conditions. The conditionsmay be acidic or neutral conditions, for example in the presence of anacid like trifluoromethanesulfonic acid, camphorsulfonic acid,p-toluenesulfonic acid or their respective pyridinium or substitutedpyridinium salts when X is CCl₃C(NH)O or in the presence of a base likepyridine, a substituted pyridine, diisopropylethylamine orpentamethylpiperidine when X is CF₃SO₃. The most preferable one is40-O-(2-hydroxy) ethyl rapamycin, which is disclosed in WO94/09010, thedisclosure of which is incorporated herein by reference.

[0069] The tricyclic compound(II), and rapamycin and its derivatives,may be in a form of its salt, which includes conventional non-toxic andpharmaceutically acceptable salt such as the salt with inorganic ororganic bases, specifically, an alkali metal salt such as sodium saltand potassium salt, an alkali earth metal salt such as calcium salt andmagnesium salt, an ammonium salt and an amine salt such as triethylaminesalt and N-benzyl-N-methylamine salt.

[0070] With respect to the immunosuppressant usable in the presentinvention, particularly the tricyclic macrolide compounds, it is to beunderstood that there may be conformers and one or more stereoisomerssuch as optical and geometrical isomers due to asymmetric carbon atom(s)or double bond(s), and such conformers and isomers are also includedwithin the scope of the present invention. And further, the tricyclicmacrolide compounds can be in the form of a solvate, which is includedwithin the scope of the present invention. The solvate preferablyinclude a hydrate and an ethanolate.

[0071] Further example of the immunosuppressant is cyclosporin and itsderivatives such as cyclosporin A, B, C, D, E, F, G, etc, which areshown in THE MERCK INDEX (12th edition), No. 2821, U.S. Pat. Nos.4,117,118, 4,215,199, 4,288,431, 4,388,307, Helv. Chim. Acta. 60,1568(1977) and 65, 1655(1982), Transplant. Proc. 17, 1362(1985), and soon. Among which, the most preferable one is cyclosporin A. Thedisclosures of the above references are incorporated herein.

[0072] In the present invention, the lipopeptide compound is shown bythe following formula(I).

[0073] Wherein R¹ is acyl group,

[0074] R² is hydrogen or hydroxy and

[0075] R³ is hydrogen or hydroxy,

[0076] or a salt thereof.

[0077] Suitable salt of the lipopeptide compound(I) is apharmaceutically acceptable and conventional non-toxic salt, and mayinclude a salt with a base or an acid addition salt such as a salt withan inorganic base, for example, an alkali metal salt (e.g., sodium salt,potassium salt, etc.), an alkaline earth metal salt (e.g., calcium salt,magnesium salt, etc.), an ammonium salt; a salt with an organic base,for example, an organic amine salt (e.g., triethylamine salt,diisopropylethylamine salt, pyridine salt, picoline salt, ethanolaminesalt, triethanolamine salt, dicyclohexylamine salt,N,N′-dibenzylethylenediamine salt, etc.); an inorganic acid additionsalt (e.g., hydrochloride hydrobromide, sulfate, phosphate, etc.);

[0078] an organic carboxylic sulfonic acid addition salt (e.g., formate,acetate, trifluoroacetate, maleate, tartrate, fumarate,methanesulfonate, benzenesulfonate, toluenesulfonate, etc.); a salt witha basic or acidic amino acid (e.g., arginine, aspartic acid, glutamicacid, etc.).

[0079] It is to be noted that each of the lipopeptide compound(I) mayinclude one or more stereoisomer(s) such as optical isomer(s) andgeometrical isomer(s) due to asymmetric carbon atom(s) and doublebond(s), and all such isomer(s) and the mixture thereof are includedwithin the scope of the present invention.

[0080] The lipopeptide compound(I) or a salt thereof includes solvatedcompound [e.g., hydrate, ethanolate, etc.)].

[0081] The lipopeptide compound(I) or a salt thereof includes both itscrystal form and non-crystal form.

[0082] It should be understood that the lipopeptide compound(I) in thepresent invention may include the prodrug form.

[0083] In the lipopeptide compound(I), suitable example of “acyl group”may include aliphatic acyl, aromatic acyl, arylaliphatic acyl andheterocyclic-aliphatic acyl derived from carboxylic acid, carbonic acid,carbamic acid, sulfonic acid, and the like.

[0084] Suitable example of said “acyl group” may be illustrated asfollows.

[0085] Aliphatic acyl such as lower or higher alkanoyl (e.g., formyl,acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl,2,2-dimethylpropanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl,decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl,pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl,icosanoyl, etc.);

[0086] lower or higher alkoxycarbonyl (e.g., methoxycarbonyl,ethoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbonyl,heptyloxycarbonyl, etc.);

[0087] lower or higher alkylsulfonyl (e.g., methylsulfonyl,ethylsulfonyl, etc.);

[0088] lower or higher alkoxysulfonyl (e.g., methoxysulfonyl,ethoxysulfonyl, etc.); or the like;

[0089] Aromatic acyl such as

[0090] aroyl (e.g., benzoyl, toluoyl, naphthoyl, etc.); aroyl which hasone or more suitable substituent(s); ar(lower)alkanoyl [e.g.,phenyl(C₁-C₆)alkanoyl (e.g., phenylacetyl, phenylpropanoyl,phenylbutanoyl, phenylisobutanoyl, phenylpentanoyl, phenylhexanoyl,etc.), naphthyl(C₁-C₆)alkanoyl (e.g., naphthylacetyl, naphthylpropanoyl,naphthylbutanoyl, etc.), etc.];

[0091] ar(lower)alkenoyl [e.g., phenyl(C₃-C₆)alkenoyl (e.g.,phenylpropenoyl, phenylbutenoyl, phenylmethacryloyl, phenylpentanoyl,phenylhexenoyl, etc.), naphthyl(C₃-C₆)alkenoyl (e.g., naphthylpropenoyl,naphthylbutenoyl, etc.), etc.];

[0092] ar(lower)alkoxycarbonyl [e.g., phenyl(C₁-C₆)alkoxycarbonyl (e.g.,benzyloxycarbonyl, etc.), fluorenyl(C₁-C₆)alkoxy-carbonyl (e.g.,fluorenylmethyloxycarbonyl, etc.), etc.];

[0093] aryloxycarbonyl (e.g., phenoxycarbonyl, naphthyloxycarbonyl,etc.);

[0094] aryloxy(lower)alkanoyl (e.g., phenoxyacetyl, phenoxypropionyl,etc.);

[0095] arylcarbamoyl (e.g., phenylcarbamoyl, etc.);

[0096] arylthiocarbamoyl (e.g., phenylthiocarbamoyl, etc.);

[0097] arylglyoxyloyl (e.g., phenylglyoxyloyl, naphthylglyoxyloyl,etc.);

[0098] arylsulfonyl which may have 1 to 4 lower alkyl (e.g.,phenylsulfonyl, p-tolylsulfonyl, etc.); or the like;

[0099] Heterocyclic acyl such as

[0100] heterocycliccarbonyl; heterocyclic(lower)alkanoyl (e.g.,heterocyclicacetyl, heterocyclicpropanoyl, heterocyclicbutanoyl,heterocyclicpentanoyl, heterocyclichexanoyl, etc.);

[0101] heterocyclic(lower)alkenoyl (e.g., heterocyclicpropenoyl,heterocyclicbutenoyl, heterocyclicpentenoyl, heterocyclichexenoyl,etc.);

[0102] heterocyclicglyoxyloyl; or the like.

[0103] Among them, more preferred “acyl group” is aroyl which has one ormore suitable substituent(s).

[0104] Suitable example of “suitable substituent(s)” in the term of“aroyl which has one or more suitable substituent(s)” may beheterocyclic group substituted with aryl having lower alkoxy,heterocyclic group substituted with aryl having loweralkoxy(lower)alkoxy, heterocyclic group substituted with aryl havinglower alkoxy(higher)alkoxy, heterocyclic group substituted with arylhaving cyclo(lower)alkyloxy, heterocyclic group substituted with arylhaving heterocyclic group, heterocyclic group substituted withcyclo(lower)alkyl having cyclo(lower)alkyl, heterocyclic groupsubstituted with aryl having aryl substituted with loweralkoxy(lower)alkoxy, heterocyclic group substituted with aryl havingheterocyclic group substituted with cyclo(lower)alkyl;

[0105] in which the preferred one may be unsaturated 3 to 8-memberedheteromonocyclic group containing 1 to 2 oxygen atom(s) and 1 to 3nitrogen atom(s) substituted with phenyl having (C₄-C₆)alkoxy,unsaturated condensed heterocyclic group containing 1 or 2 sulfuratom(s) and 1 to 3 nitrogen atom(s) substituted with phenyl having(C₄-C₆)alkoxy, unsaturated 3 to 8-membered heteromonocyclic groupcontaining 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s) substitutedwith phenyl having (C₁-C₄)alkoxy(C₄-C₆)alkoxy, unsaturated 3 to8-membered heteromonocyclic group containing 1 or 2 sulfur atom(s) and 1to 3 nitrogen atom(s) substituted with phenyl having(C₁-C₄)alkoxy(C₇-C₁₄)alkoxy, saturated 3 to 8-membered heteromonocyclicgroup containing 1 to 4 nitrogen atom(s) substituted with phenyl having(C₁-C₄)alkoxy(C₇-C₁₄)alkoxy, unsaturated condensed heterocyclic groupcontaining 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s) substitutedwith phenyl having cyclo(C₄-C₆)alkyloxy, unsaturated condensedheterocyclic group containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogenatom(s) substituted with phenyl saturated 3 to 8-memberedheteromonocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3nitrogen atom(s), saturated 3 to 8-membered heteromonocyclic groupcontaining 1 to 4 nitrogen atom(s) substituted with cyclo(C₄-C₆)alkylhaving cyclo(C₄-C₆)alkyl, unsaturated 3 to 8-membered heteromonocyclicgroup containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s)substituted with phenyl having phenyl substituted with(C₁-C₄)alkoxy(C₁-C₄)alkoxy, unsaturated 3 to 8-membered heteromonocyclicgroup containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s)substituted with phenyl having saturated 3 to 8-memberedheteromonocyclic group containing 1 to 4 nitrogen atom(s) substitutedwith cyclo(C₄-C₆)alkyl, unsaturated condensed heterocyclic groupcontaining 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s) substitutedwith phenyl having saturated 3 to 8-membered heteromonocyclic groupcontaining 1 to 4 nitrogen atom(s) having cyclo(C₄-C₆)alkyl, etc.

[0106] Among them, the most preferred one may be isoxazolyl substitutedwith phenyl having pentyloxy, imidazothiadiazolyl substituted withphenyl having pentyloxy, thiadiazolyl substituted with phenyl havingmethoxyhexyloxy, thiadiazolyl substituted with phenyl havingmethoxyoctyloxy, thiadiazolyl substituted with phenyl havingmethoxyheptyloxy, imidazothiadiazolyl substituted with phenyl havingcyclohexyloxy, imidazothiadiazolyl substituted with phenyl havingdimethylmorpholino, piperazinyl substituted with phenyl havingmethoxyheptyloxy, piperazinyl substituted with phenyl havingmethoxyoctyloxy, piperazinyl substituted with cyclohexyl havingcyclohexyl, thiadiazolyl substituted with phenyl having phenylsubstituted with methoxyethoxy, thiadiazolyl substituted with phenylhaving phenyl substituted with methoxybutoxy, thiadiazolyl substitutedwith phenyl having phenyl substituted with ethoxypropoxy,imidazothiadiazolyl substituted with phenyl having piperazinylsubstituted with cyclohexyl, imidazothiadiazolyl substituted with phenylhaving piperazinyl substituted with cyclohexyl.

[0107] The more suitable example of “acyl group” of R¹ may be benzoylwhich has isoxazolyl substituted with phenyl having pentyloxy, benzoylwhich has imidazolthiadiazolyl substituted with phenyl having pentyloxy,benzoyl which has thiadiazolyl substituted with phenyl havingmethoxyhexyloxy, benzoyl which has thiadiazolyl substituted with phenylhaving methoxyoctyloxy, benzoyl which has thiadiazolyl substituted withphenyl having methoxyheptyloxy, benzoyl which has imidazothiadiazolylsubstituted with phenyl having cyclohexyloxy, benzoyl which hasimidazothiadiazolyl substituted with phenyl having dimethylmorpholino,benzoyl which has piperazinyl substituted with phenyl havingmethoxyheptyloxy, benzoyl which has piperazinyl substituted with phenylhaving methoxyoctyloxy, benzoyl which has piperazinyl substituted withcyclohexyl having cyclohexyl, benzoyl which has thiadiazolyl substitutedwith phenyl having phenyl substituted with methoxyethoxy, benzoyl whichhas thiadiazolyl substituted with phenyl having phenyl substituted withmethoxybutoxy, benzoyl which has thiadiazolyl substituted with phenylhaving phenyl substituted with ethoxypropoxy, benzoyl which hasimidazothiadiazolyl substituted with phenyl having piperazinylsubstituted with cyclohexyl, benzoyl which has imidazothiadiazolylsubstituted with phenyl having piperazinyl substituted with cyclohexyl.

[0108] In the present invention, an immunosuppressant is able to beadministered for increasing an antifungal activity of the lipopeptidecompound(I) simultaneously, separately or in sequential use with thelipopeptide compound(I).

[0109] If advisable, the lipopeptide compound(I) can be mixed with theimmunosuppressant prior to its use. So, the composition comprising thesaid the immunosuppressant of the present invention may further comprisethe lipopeptide compound(I). And optionally, it comprises furtheradditional ingredients, such as, mycophenolate mofetil (CellCept),steroids, Azathiopurine, and so on.

[0110] While the effective dosage of the immunosuppressant depends onthe type of the said immunosuppressant, the patient's age, type ofdisease, severity of illness, and other factors, a daily dose thereof isabout 0.01˜1000 mg, preferably 0.05˜500 mg, and more preferably, 0.1˜100mg for therapeutic purposes. The average unit dose may be generallyabout 0.1 mg, 0.5 mg, 1 mg, 5 mg, 10 mg, 50 mg, 100 mg, 250 mg, or 500mg.

[0111] The pharmaceutical composition of the immunosuppressant can beused in the form of a pharmaceutical preparation, for example, in solid,semisolid or liquid form, which contains the immunosuppressant or apharmaceutically acceptable salt thereof, as an active ingredient inadmixture with an organic or inorganic carrier or excipient which issuitable for rectal; pulmonary (nasal or buccal inhalation); ocular;external (topical); oral administration; parenteral (includingsubcutaneous, intravenous and intramuscular) administrations;insufflation (including aerosols from metered dose inhalator);nebulizer; or dry powder inhalator.

[0112] The active ingredient may be compounded, for example, with theusual non-toxic, pharmaceutically acceptable carriers in a solid formsuch as granules, tablets, dragees, pellets, troches, capsules, orsuppositories; creams; ointments; aerosols; powders for insufflation; ina liquid form such as solutions, emulsions, or suspensions forinjection; ingestion; eye drops; and any other form suitable for use.And, if necessary, there may be included in the above preparationauxiliary substance such as stabilizing, thickening, wetting,emulsifying and coloring agents; perfumes or buffer; or any othercommonly may be used as additives.

[0113] The lipopeptide compound(I) or a pharmaceutically acceptable saltthereof may also be included in the pharmaceutical composition of thepresent invention in an amount sufficient to produce the desiredantimicrobial effect upon the process or condition of diseases.

[0114] For applying the lipopeptide compound(I) or a salt thereof tohuman, it is preferable to apply it by intravenous, intramuscular,pulmonary, oral administration, or insufflation. While the dosage oftherapeutically effective amount of the lipopeptide compound(I) variesfrom and also depends upon the age and condition of each individualpatient to be treated, in the case of intravenous administration, adaily dose of 0.01-20 mg of the lipopeptide compound(I) per kg weight ofhuman being in the case of intramuscular administration, a daily dose of0.1-20 mg of the lipopeptide compound(I) per kg weight of human being,in case of oral administration, a daily dose of 0.5-50 mg of thelipopeptide compound(I) per kg weight of human being is generally givenfor treating or preventing infectious diseases.

[0115] Especially in case of the treatment of prevention of Pneumocystiscarinii infection, the followings are to be noted.

[0116] For administration by inhalation, the lipopeptide compound(I) isconveniently delivered in the form of an aerosol spray presentation frompressurized as powders which may be formulated and the powdercompositions may be inhaled with the aid of an insufflation powderinhaler device. The preferred delivery system for inhalation is ametered dose inhalation aerosol, which may be formulated as a suspensionor solution of compound in suitable propellants such as fluorocarbons orhydrocarbons.

[0117] Because of desirability to directly treat lung and bronchi,aerosol administration is a preferred method of administration.Insufflation is also a desirable method, especially where infection mayhave spread to ears and other body cavities.

[0118] Alternatively, parenteral administration may be employed usingdrip intravenous administration.

[0119] The lipopeptide compound(I) or a salt thereof has antifungalactivities, in which the fungi may include Acremonium;

[0120] Absidia (e.g., Absidia corymbifera, etc);

[0121] Aspergillus (e.g., Aspergillus clavatus, Aspergillus flavus,Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger,Aspergillus terreus, Aspergillus versicolor, etc);

[0122] Blastomyces (e.g., Blastomyces dermatitidis, etc);

[0123] Candida (e.g., Candida albicans, Candida glabrata, Candidaguilliermondii, Candida kefyr, Candida krusei, Candida parapsilosis,Candida stellatoidea, Candida tropicalis, Candida utilis, etc.);

[0124] Cladosporium (e.g., Cladosporium trichoides, etc);

[0125] Coccidioides (e.g., Coccidioides immitis, etc);

[0126] Cryptococcus (e.g., Cryptococcus neoformans, etc);

[0127] Cunninghamella (e.g., Cunninghamella elegans, etc);

[0128] Dermatophyte;

[0129] Exophiala (e.g., Exophiala dermatitidis, Exophiala spinifera,etc);

[0130] Epidermophyton (e.g., Epidermophyton floccosum, etc);

[0131] Fonsecaea (e.g., Fonsecaea pedrosoi, etc);

[0132] Fusarium (e.g., Fusarium solani, etc);

[0133] Geotrichum (e.g., Geotrichum candiddum, etc);

[0134] Histoplasma (e.g., Histoplasma capsulatum var. capsulatum, etc);

[0135] Malassezia (e.g., Malassezia furfur, etc);

[0136] Microsporum (e.g., Microsporum canis, Microsporum gypseum, etc);

[0137] Mucor;

[0138] Paracoccidioides (e.g., Paracoccidioides brasiliensis, etc);

[0139] Penicillium (e.g., Penicillium marneffei, etc);

[0140] Phialophora;

[0141] Pneumocystis (e.g., Pneumocystis carinii, etc);

[0142] Pseudallescheria (e.g., Pseudallescheria boydii, etc);

[0143] Rhizopus (e.g., Rhizopus microsporus var. rhizopodiformis,Rhizopus oryzae, etc);

[0144] Saccharomyces (e.g., Saccharomyces cerevisiae, etc);

[0145] Scopulariopsis;

[0146] Sporothrix (e.g., Sporothrix schenckii, etc);

[0147] Trichophyton (e.g., Trichophyton mentagrophytes, Trichophytonrubrum, etc);

[0148] Trichosporon (e.g., Trichosporon asahii, Trichosporon cutaneum,etc).

[0149] The above fungi are well known to cause various infectiondiseases in skin, hair, nail, oral mucosa, gastrointestinal tract,bronchus, lung, endocardium, brain, meninges, urinary organ, vaginalprotion, oral cavity, ophthalmus, systemic, kidney, bronchus, heart,external auditory canal, bone, nasal cavity, paranasal cavity, spleen,liver, hypodermal tissue, lymph duct, gastrointestine, articulation,muscle, tendon, interstitial plasma cell in lung, and so on.

[0150] Therefore, the composition of the present invention are usefulfor preventing and treating various infectious diseases, such asdermatophytosis(e.g., trichophytosis, etc), pityriasis versicolor,candidiasis, cryptococcosis, geotrichosis, trichosporosis,aspergillosis, penicilliosis, fusariosis, zygomycosis, sporotrichosis,chromomycosis, coccidioidomycosis, histoplasmosis, blastomycosis,paracoccidioidomycosis, pseudallescheriosis, mycetoma, mycotickeratitis, otomycosis, pneumocystosis, and so on.

[0151] Particularly, the lipopeptide compound(I) or a salt thereof hasan inhibitory activity on β-1,3-glucan synthase, and therefore which areexpected to be useful for the prophylactic and/or therapeutic treatmentof Pneumocystis carinii infection (e.g. Pneumocystis carinii pneumonia)in a human being or an animal (WO96/11210 and WO99/40108). Therefore,the present composition is useful for prophylactic and/or therapeutictreatment of infectious diseases caused by the above-mentioned fungi.

[0152] From the another aspect, the present invention also provides thefollowing inventions.

[0153] i) A use of an immunosuppressant for manufacturing a medicamentfor increasing an effect caused by a lipopeptide compound, in which theeffect caused by a lipopeptide compound is an antifungal activity.

[0154] ii) A use of an immunosuppressant and a lipopeptide compound formanufacturing a medicament for simultaneous, separate or sequential usefor an antifungal activity.

[0155] iii) A method for increasing an antifungal activity of alipopeptide compound, by administering an effective amount ofimmunosuppressant to a human being or an animal.

[0156] iv) A method for preventing and treating infectious diseases, byadministering an effective amount of a lipopeptide compound and aneffective amount of an immunosuppressant to a human being or an animal.

[0157] v) A composition comprising an immunosuppressant for increasingan antifungal activity of a lipopeptide compound.

[0158] vi) A composition comprising an immunosuppressant and alipopeptide compound as a combined preparation for simultaneous,separate or sequential use for an antifungal activity.

[0159] vii) An article of manufacture, comprising packaging material andthe composition mentioned in the above v) or vi) contained within saidthe composition is therapeutically effective for infectious diseases,and wherein said packaging material comprises a label or a writtenmaterial which indicates that said composition can be used forinfectious diseases.

[0160] viii) A commercial package comprising the composition mentionedin the above v) or vi), and a written matter associated therewith,wherein the written matter states that the composition can or should beused for preventing or treating infectious diseases.

[0161] The invention is further illustrated in connection with thefollowing non-limiting example.

[0162] Test Compounds

[0163] Test Method

[0164] The broth microdilloution method using RPMI medium (pH7.0) wasused, comparing the each drug alone (Test Compound (A) and Test Compound(B)) and combined for clinical isolate of Aspergillus fumigatus. Acombination of drug concentrations was evaluated by the checkerboaredmethod.

[0165] All wells were examined macroscopically for growth and comparedto a control (no drug). MIC was visually determined as the lowestconcentration resulting in prominent decrease in turbidity compared tocontrols.

[0166] The Fractional Inhibitory Concentration(FIC) for each drug inmixture wells was compared to the MIC for each drug alone. The FIC indexwas calculated from the sum of the FICs for the two drugs. Aquantitative expression of the interaction for inhibition is as follows:

Synergy=<0.5;

[0167] Test Result

[0168] In vitro combination with Test Compound (A) and Test Compound (B)against A. fumigatus MIC (μg/ml) Test Test Test Com- Test Com- Com-pound Com- pound (B) pound (A) Com- pound (B) com- Organism Alonebination Alone bination FIC index A. fumigatus 0.0156 0.0039 0.1250.0313 0.50 FP1996

[0169] From the results of the above example, synergy effect of efficacywas observed with combination of the lipopeptide compound(I) and theimmunosuppressant at certain concentrations. No antagonism of efficacywith the immunosuppressant in combination with the lipopeptidecompound(I) also was seen.

[0170] Given the above disclosure, it is thought that variations willoccur to those skilled in the art. For example, it is thought thatcombination using other immunosuppressant and the lipopeptide compoundsother than the lipopeptide compound(I) may also be effective againstfungal infections caused by the fungal pathogens noted. Accordingly, itis intended that the above example should be construed as illustrativeand that the invention disclosed herein should be limited only by thefollowing claims.

[0171] The patents, patent applications and publications cited hereinare incorporated by reference.

1. A use of an immunosuppressant for manufacturing a medicament forincreasing an antifungal activity of a lipopeptide compound.
 2. The useof the claim 1, in which the lipopeptide compound inhibits theproduction of β-1,3-glucan.
 3. The use of the claim 1, in which thelipopeptide compound inhibits the activity of β-1,3-glucan synthase. 4.The use of the claim 1, in which the immunosuppressant is tacrolimus orits hydrate, or cyclosporins, and the lipopeptide compound is shown bythe following formula(I):

Wherein R¹ is acyl group, R² is hydrogen or hydroxy and R³ is hydrogenor hydroxy, or a salt thereof.
 5. A use of an immunosuppressant and alipopeptide compound for manufacturing a medicament for simultaneous,separate or sequential use for an antifungal activity.
 6. A method forincreasing an antifungal activity of a lipopeptide compound, byadministering an effective amount of an immunosuppressant to a humanbeing or an animal.
 7. A method for preventing and treating infectiousdiseases, by administering an effective amount of an immunosuppressantand an effective amount of a lipopeptide compound to a human being or ananimal.
 8. A composition comprising an immunosuppressant, for increasingan antifungal activity of a lipopeptide compound.
 9. A compositioncomprising an immunosuppressant and a lipopeptide compound as a combinedpreparation for simultaneous, separate or sequential use for anantifungal activity.
 10. An article of manufacture, comprising packagingmaterial and the composition claimed in claim 8 or 9 contained withinsaid packaging material, wherein said composition is therapeuticallyeffective for infectious diseases, and wherein said packaging materialcomprises a label or a written material which indicates that saidcomposition can be used for infectious diseases.
 11. A commercialpackage comprising the composition of any of claims 8 and 9, and awritten matter associated therewith, wherein the written matter statesthat the composition can or should be used for preventing or treatinginfectious diseases.